CKIα plus CaMKII double knockdown reduced NS5A S235 phosphorylation and HCV RNA levels in the infected Huh7.5.1 cells.

<p>(A) Viability of the Huh7.5.1 cells with CKIα plus CaMKII double knockdown. (B) Representative and summary (C) of the immunoblotting for NS5A and NS5A phosphorylation at S235 (pS235) in the HCV-infected Huh7.5.1 cells with or without CKIα plus CaMKII double knockdown. One day after the cells were infected with HCV (J6/JFH1), the cells were subjected to shRNA-based double knockdown for 6 days prior to the immunoblotting analysis. Values are Mean ± SEM (n = 3). Asterisk indicates significance i.e. p<0.05, t-test against the values in the non-targeted control cells (shCon). Protein abundance was quantified with the Li‐Cor scanner, adjusted for the loadings (i.e. against β-actin) and normalized with the values in the control cells. (D and E) Quantitative measurements of kinase mRNA and HCV RNA in the HCV-infected Huh7.5.1 cells with or without CKIα plus CaMKII double knockdown. The experiments were done as those in b except that RNA was collected for the analysis. The GAPDH mRNA levels were analyzed as loading controls. Relative RNA abundance was normalized to the values of the control cells.